Eficacia y seguridad de L-asparaginasa erwinia y L-asparaginasa E. Coli pegilada en el tratamiento de pacientes con leucemia linfoblástica aguda que presentam hipersensibilidad a L-asparaginasa E. coli nativa / Efficacy and safety of L-asparaginase erwinia and L-asparaginase E. coli pegylated in the treatment of patients with acute lymphoblastic leukemia who are hypersensitive to L-asparaginase E. coli native

INTRODUÇÃO: Antecedentes: El presente dictamen expone la evaluación de tecnología de la eficacia y seguridad del uso de L-asparaginasa Erwinia y L-asparaginasa E. coli pegilada para el tratamiento de pacientes con leucemia linfoblástica aguda que presentan hipersensibilidad a L-asparaginasa E. coli nativa. Aspectos Generales: La leucemia es el tipo de cáncer más común en niños, representando aproximadamente el 30% de todos los tipos de cáncer diagnosticados en niños; siendo la leucemia linfoblástica aguda (LLA) uno de los dos tipos de leucemias más comunes. Adicionalmente, alrededor del 60% de todos los casos de LLA ocurre en pacientes menores de 20 años. Así, LLA es un tipo de leucemia de alta importancia dentro de población joven. Tecnología Sanitaria de Interés: Las células neoplásicas en la leucemia linfoblástica aguda (LLA) no sintetizan las cantidades necesarias del aminoácido L-asparagina; por lo que requieren de funtes externas (i.e., L-asparagina extracelular). La L-asparaginasa, es una enzima que cataliza la conversión de L-asparagina más agua, en ácido aspártico y amoniaco, ocasionando que los niveles de L-asparagina extracelular disminuyan; y que por los tanto las células d ela LLA no cuenten con L-asparagina extracelular. Así, estas células neoplásicas se quedan sin fuentes de L-asparagina, y no pueden sintetizar proteínas de gran imporancia para su supervivencia, ocasionando su muerte. METODOLOGÍA: Estrategia de Búsqueda: Se realizó una búsqueda de la literatura a la eficacia y seguridad del uso de L-asparaginasa Erwinia y L-asparaginasa E. coli pegilada para el tratamiento de pacientes niños y adultos con leucemia linfoblástica aguda que presentan hipersensibilidad a L-asparaginasa E. coli nativa. Esta búsqueda se realizó utilizando los meta-buscadores: Translating Research into Practice (TRIPDATABASE) Y National Library of Medicine (Pubmed-Medline). RESULTADOS: Sinopsis de la Evidencia: Se realizó la búsqueda bibliográfica y de evidencia científica hasta marzo del 2017 para el sustento del uso de L-asparaginasa Erwinia en el tratamiento de leucemia linfoblástica aguda en pacientes niños y adultos que presentan hipersensibilidad a L-asparaginasa E. coli nativa. Se presente la evidencia disponible según el tipo de publicación priorizada en los criterios de inclusión (i.e., GP, ETS, RS y ECA fase III), siendo los ensayos de fase III o en su defecto ensayos controlados y aleatorizados la principal considerada. CONCLUSIONES: El presente dictamen evaluó la mejor evidencia disponible hasta marco 2017 en relación al uso de L-asparaginasa Erwinia y L-asparaginasa E. coli pegilada para el tratamiento de pacientes niños, adolescentes, y adultos con leucemia linfoblástica aguda, que presentan hipersensibilidad a L-asparaginasa E. coli nativa. El Instituto de Tecnologías Sanitarias-IETSI, aprueba el uso de L-asparaginasa Erwinia como parte del esquema quimioterápico utilizada para el tratamiento de pacientes niños, adolescentes, y adultos con leucemia linfoblástica aguda, que presentan hipersensibilidad grado 2 o más a L-asparaginasa E. coli nativa. La vigencia de este dictamen preliminar es de dos años a partir de la fecha de publicación. Asimismo, el Instituto de Tecnologías Sanitarias-IETSI, no aprueba el uso de L-asparaginasa E. coli pegilada en el tratamiento de pacientes niños, adolescentes, y adultos con leucemia linfoblástica aguda, que presentan hipersensiblidad grado 2 o más a L-asparaginasa E. coli nativa.

Peptidoglycan sensing by the receptor PGRP-LE in the Drosophila gut induces immune responses to infectious bacteria and tolerance to microbiota.

Gut epithelial cells contact both commensal and pathogenic bacteria, and proper responses to these bacteria require a balance of positive and negative regulatory signals. In the Drosophila intestine, peptidoglycan-recognition proteins (PGRPs), including PGRP-LE, play central roles in bacterial recognition and activation of immune responses, including induction of the IMD-NF-κB pathway. We show that bacteria recognition is regionalized in the Drosophila gut with various functional regions requiring different PGRPs. Specifically, peptidoglycan recognition by PGRP-LE in the gut induces NF-κB-dependent responses to infectious bacteria but also immune tolerance to microbiota through upregulation of pirk and PGRP-LB, which negatively regulate IMD pathway activation. Loss of PGRP-LE-mediated detection of bacteria in the gut results in systemic immune activation, which can be rescued by overexpressing PGRP-LB in the gut. Together these data indicate that PGRP-LE functions as a master gut bacterial sensor that induces balanced responses to infectious bacteria and tolerance to microbiota.

Comprehending crystalline ß-carotene accumulation by comparing engineered cell models and the natural carotenoid-rich system of citrus.

Genetic manipulation of carotenoid biosynthesis has become a recent focus for the alleviation of vitamin A deficiency. However, the genetically modified phenotypes often challenge the expectation, suggesting the incomplete comprehension of carotenogenesis. Here, embryogenic calli were engineered from four citrus genotypes as engineered cell models (ECMs) by over-expressing a bacterial phytoene synthase gene (CrtB). Ripe flavedos (the coloured outer layer of citrus fruits), which exhibit diverse natural carotenoid patterns, were offered as a comparative system to the ECMs. In the ECMs, carotenoid patterns showed diversity depending on the genotypes and produced additional carotenoids, such as lycopene, that were absent from the wild-type lines. Especially in the ECMs from dark-grown culture, there emerged a favoured ß,ß-pathway characterized by a striking accumulation of ß-carotene, which was dramatically different from those in the wild-type calli and ripe flavedos. Unlike flavedos that contained a typical chromoplast development, the ECMs sequestered most carotenoids in the amyloplasts in crystal form, which led the amyloplast morphology to show a chromoplast-like profile. Transcriptional analysis revealed a markedly flavedo-specific expression of the ß-carotene hydroxylase gene (HYD), which was suppressed in the calli. Co-expression of CrtB and HYD in the ECMs confirmed that HYD predominantly mediated the preferred carotenoid patterns between the ECMs and flavedos, and also revealed that the carotenoid crystals in the ECMs were mainly composed of ß-carotene. In addition, a model is proposed to interpret the common appearance of a favoured ß,ß-pathway and the likelihood of carotenoid degradation potentially mediated by photo-oxidation and vacuolar phagocytosis in the ECMs is discussed.

Hypersensitive response and acyl-homoserine lactone production of the fire blight antagonists Erwinia tasmaniensis and Erwinia billingiae.

Fire blight caused by the Gram-negative bacterium Erwinia amylovora can be controlled by antagonistic microorganisms. We characterized epiphytic bacteria isolated from healthy apple and pear trees in Australia, named Erwinia tasmaniensis, and the epiphytic bacterium Erwinia billingiae from England for physiological properties, interaction with plants and interference with growth of E. amylovora. They reduced symptom formation by the fire blight pathogen on immature pears and the colonization of apple flowers. In contrast to E. billingiae, E. tasmaniensis strains induced a hypersensitive response in tobacco leaves and synthesized levan in the presence of sucrose. With consensus primers deduced from lsc as well as hrpL, hrcC and hrcR of the hrp region of E. amylovora and of related bacteria, these genes were successfully amplified from E. tasmaniensis DNA and alignment of the encoded proteins to other Erwinia species supported a role for environmental fitness of the epiphytic bacterium. Unlike E. tasmaniensis, the epiphytic bacterium E. billingiae produced an acyl-homoserine lactone for bacterial cell-to-cell communication. Their competition with the growth of E. amylovora may be involved in controlling fire blight.

Evaluation of immunologic crossreaction of antiasparaginase antibodies in acute lymphoblastic leukemia (ALL) and lymphoma patients.

To evaluate how well antibodies to one asparaginase preparation predict or correlate with antibodies to another preparation in acute lymphoblastic leukemia (ALL) and lymphoma patients who did and did not have hypersensitivity reactions during chemotherapy. In all, 24 children with newly diagnosed ALL or lymphoma, who received Escherichia coli asparaginase 10 000 IU/m(2) IM thrice weekly for nine doses as part of multiagent induction and reinduction chemotherapy, and seven monthly doses during the first 7 months of continuation treatment, were studied. Plasma samples were collected at postinduction and at postreinduction. Six of 24 patients had no overt clinical reactions (nonreacting) and received only the E. coli preparation. Of these, 18 patients who had allergic reactions were switched to Erwinia asparaginase. A total of 18 patients had an anaphylactoid reaction to Erwinia asparaginase and were switched to receive polyethylene glycol (PEG) asparaginase. Antibody levels were measured by enzyme-linked immunoadsorbent assay against all the three asparaginase preparations. At postinduction, antibodies against E. coli were higher in reacting patients (0.063+/-0.066) than in nonreacting patients (0.019+/-0.013) (P=0.03). At postreinduction, anti-Erwinia antibodies were significantly higher in reacting patients (0.431+/-0.727) than in nonreacting patients (0.018+/-0.009) (P=0.007). Anti-E. coli antibodies correlated with anti-PEG antibodies at postinduction (r=0.714, P<0.001) and at postreinduction (r=0.914, P<0.001), but did not correlate with anti-Erwinia antibodies at postinduction (r=0.119, P=0.580) and at postreinduction (r=0.078, P=0.716). The results indicate a crossreactivity between patient antibodies raised against natural E. coli and PEG asparaginase but not Erwinia asparaginase.

Anti-Erwinia asparaginase antibodies during treatment of childhood acute lymphoblastic leukemia and their relationship to outcome: a case-control study.

PURPOSE: A case-control study was performed to determine whether patients who had been treated with Erwinia asparaginase as part of their treatment for childhood acute lymphoblastic leukemia (ALL) and who showed relapsed of their disease more often developed anti-asparaginase antibodies than patients who remained in remission. METHODS: A group of 13 patients who showed relapsed of their disease (median follow-up 35 months) were randomly matched with control patients of the same risk group (two control patients to each case), who had received therapy of the same intensity during the same period (median follow-up 70 months). Anti- Erwinia asparaginase antibodies were measured (ELISA method) during maintenance therapy after asparaginase treatment (30,000 IU/m(2) daily for 10 days in all patients plus twice weekly for 2 weeks in intermediate-risk and high-risk ALL patients). RESULTS: The overall incidence of anti- Erwinia asparaginase antibodies was 8% (3 of 39 patients). There was no statistically significant difference in the incidence of antibody formation between patients who had suffered relapse (1 of 13) and those who had not (2 of 26). In two of the three patients who developed antibodies, the antibodies disappeared after some time, whereas one patient had measurable antibody levels for more than a year after asparaginase therapy. CONCLUSIONS: In this study, the development of anti-Erwinia asparaginase antibodies was rare and was unrelated to the risk of relapse.

Mutation in a gene required for lipopolysaccharide and enterobacterial common antigen biosynthesis affects virulence in the plant pathogen Erwinia carotovora subsp. atroseptica.

Spontaneous bacteriophage-resistant mutants of the phytopathogen Erwinia carotovora subsp. atroseptica (Eca) SCRI1043 were isolated and, out of 40, two were found to exhibit reduced virulence in planta. One of these mutants, A5/22, showed multiple cell surface defects including alterations in synthesis of outer membrane proteins, lipopolysaccharide (LPS), enterobacterial common antigen (ECA), and flagella. Mutant A5/22 also showed reduced synthesis of the exoenzymes pectate lyase (Pel) and cellulase (Cel), major virulence factors for this pathogen. Genetic analysis revealed the pronounced pleiotropic mutant phenotype to be due to a defect in a single gene (rffG) that, in Escherichia coli, is involved in the production of ECA. We also show that while other enteric bacteria possess duplicate homologues of this gene dedicated separately to synthesis of LPS and ECA, Eca has a single gene.

Effect of inhalation of organic dust-derived microbial agents on the pulmonary phagocytic oxidative metabolism of guinea pigs.

The effect of inhalation exposure of various biological agents associated with organic dusts on the function of guinea pigs pulmonary phagocytes was investigated. Agents included antigens of Erwinia herbicola, Thermoactinomyces vulgaris, and Aspergillus fumigatus; endotoxin of Erwinia herbicola; bacterial protease; and a fungal glucan preparation. Pulmonary parameters monitored in this study were cellular differential counts from bronchoalveolar lavage, and superoxide anion and/or hydrogen peroxide production by phagocytic cells. Most of the agents caused an influx of neutrophils, lymphocytes, and red blood cells to the lung, and an enhancement of secretion of reactive oxygen species by pulmonary phagocytes. However, the relative magnitude of the inflammatory response varied greatly among these biological agents. In general, antigens of Erwinia herbicola and Aspergillus fumigatus were most potent, while bacterial protease was least effective.

Serological difference between Erwinia herbicola strains of plant and human origin.

Fifty Erwinia herbicola isolates obtained from host plants were examined in an agglutination reaction with antiserum prepared against E. ananas (E. herbicola) strain CCM 2407 antigen of plant origin and with antiserum prepared against Enterobacter agglomerans strain CNCTC M 269 antigen of human origin. In tests with strain CCM 2407 antiserum, 56% isolates showed a positive reaction, while in tests with strain CNCTC M 269 antiserum only 14% isolates showed a positive reaction. Among E. herbicola isolates which showed a positive reaction with CCM 2407 antiserum 18% showed a positive reaction with the CNCTC M 269 antiserum too. Our results confirmed the serological heterogeneity of E. herbicola population. In spite of the difference in the origin of the two antigens used for the preparation of antisera (plant, human; Japan, Czech Republic) our results indicate that some of our E. herbicola strains and E. agglomerans strain CNCTC M 269 are serologically identical.

[Pathogenic action of Erwinia herbicola. II. In vivo investigations]. / Patogenne dzialanie paleczki Erwinia herbicola. II. Badania in vivo.

The pulmonary response of guinea pigs to inhalation of antigen and endotoxin of Erwinia herbicola was investigated. Pulmonary parameters monitored in this study were breathing rate, cellular differential counts from bronchoalveolar lavage fluid, superoxide anion and interleukin-1 (IL-1) production by alveolar macrophages. Both agents caused an increase in breathing rate; an influx of neutrophils, lymphocytes, and red blood cells to the lung; and an enhancement of secretion of superoxide anions and IL-1 by pulmonary macrophages. The results indicate significant role of extrinsic bacilli Erwinia herbicola in respiratory pathology and show new mechanisms of their action.

[Results of bronchoprovocation tests with the allergen Erwinia herbicola in persons with occupational exposure to organic dust]. / Wyniki testów wziewnych z alergenem Erwinia herbicola u osób z zawodowym narazeniem na pyl organiczny.

The study was carried out on 290 males with occupational organic dust exposition. In all bronchoprovocation test with Erwinia herbicola antigen were carried out. A positive test was seen in 16.2% of the studied subjects. Bronchial hyperreactivity to Erwinia herbicola allergen depended on living environs, occupational exposure and symptoms characteristic of allergic alveolitis.

Purification and characterization of extracellular pectate lyase from Bacillus subtilis.

Bacillus subtilis strain SO113 secretes a pectate lyase which is produced during the exponential death phase of growth, just before sporulation. This extracellular pectate lyase, which produces unsaturated products from polygalacturonate, was purified 35-fold from the culture supernatant of Bacillus subtilis by a CM Sephadex chromatography. It has an isoelectric point of about 9.6 and an Mr of 42,000. Optimum activity occurred at pH 8.4 and at 42 degrees C. Calcium has a stimulative effect on the enzyme activity while EDTA leads to enzyme inactivation. The pectate lyase has a specific activity of 131 mumol of aldehyde groups per min and per mg of protein. The Km of the purified enzyme for polygalacturonic acid was 0.862 g.l-1 and the Vmax for polygalacturonic acid hydrolysis was 1.475 mumol of unsaturated products per min and per mg of protein. By using monoclonal antibodies raised against Erwinia chrysanthemi 3937 pectate lyases, it was shown that pectate lyases b and c of this strain are immunologically closely related to the Bacillus subtilis pectate lyase.

Immunological characterization of ice nucleation proteins from Pseudomonas syringae, Pseudomonas fluorescens, and Erwinia herbicola.

Antibodies were raised against the InaW protein, the product of the ice nucleation gene of Pseudomonas fluorescens MS1650, after protein isolation from an Escherichia coli clone. On Western blots (immunoblots), these antibodies recognized InaW protein and InaZ protein (the ice nucleation gene product of Pseudomonas syringae S203), produced by both E. coli clones and the source organisms. The InaZ protein appeared in P. syringae S203 during stationary phase; its appearance was correlated with the appearance of the ice nucleation-active phenotype. In contrast, the InaW protein occurred at relatively constant levels throughout the growth phases of P. fluorescens MS1650; the ice nucleation activity was also constant. Western analyses of membrane preparations of P. syringae PS31 and Erwinia herbicola MS3000 with this antibody revealed proteins which were synthesized with development of the nucleating phenotype. In these species the presence or absence of the nucleating phenotype was controlled by manipulation of culture conditions. In all nucleation-positive cultures examined, cross-reacting low-molecular-weight bands were observed; these bands appeared to be products of proteolytic degradation of ice nucleation proteins. The proteolysis pattern of InaZ protein seen on Western blots showed a periodic pattern of fragment sizes, suggesting a highly repetitive site for protease action. A periodic primary structure is predicted by the DNA sequence of the inaZ gene.

Structure of the sidechain of lipopolysaccharide from Erwinia amylovora T.

The sidechain of lipopolysaccharide from Erwinia amylovora T was composed of D-fucose, D-galactose and D-glucose in equimolar proportions. Using NMR spectroscopy, methylation analysis, mass spectrometry, Smith degradation and optical rotation data, the repeat unit was shown to have the following most probable structure: (formula; see text)

Characterization of type 1 and mannose-resistant fimbriae of Erwinia spp.

Type 1 fimbriae from Erwinia carotovora subsp. carotovora and mannose-resistant fimbriae from Erwinia rhapontici were purified and characterized. The type 1 fimbrillin had an apparent molecular weight of 16,500; that of the mannose-resistant fimbrillin was 18,000. The amino-terminal amino acid sequences of the two fimbrillins were related, but tryptic peptide maps showed significant differences between the proteins. No serological cross-reaction was found between the two fimbrial filaments, nor did they cross-react with type 1 or type 3 fimbriae purified from other enterobacterial species. Immunofluorescent staining of bacterial populations revealed that they were heterogeneous with respect to fimbriation.

Hemagglutinating activity in phytopathogenic bacteria surface compounds.

Extracellular components of plant pathogenic bacteria were obtained from their culture medium as well as from the whole cells by using NaCl 1 M, pH 6.0; 20% sucrose dissolved in 0.03 M Tris buffer, pH 8.0; or 0.05 M Na2EDTA. All the extracts from Erwinia carotovora subsp. carotovora, Xanthomonas campestris pv. campestris, Pseudomonas syringae pv. phaseolicola, Xanthomonas campestris pv. phaseoli, Pseudomonas solanacearum, and Erwinia carotovora subsp. atroseptica, were assayed for hemagglutinating activity on sheep, rabbit and chicken red blood cells (RBCs). The only active extracts were those obtained by NaCl treatment. They agglutinated sheep and rabbit erythrocytes. Extracts from E. carotovora subsp. atroseptica gave rise to the high agglutination titer on rabbit RBCs. These extracts had the lowest polysaccharide/protein ratio. E. carotovora subsp. carotovora extracts showed only a low titer (18.5 units). The agglutinating activity present in NaCl extracts of the bacteria tested was inhibited by different carbohydrates to various extent. Extracts from E. carotovora subsp. atroseptica appeared to be the most sensitive ones while those of E. carotovora subsp. carotovora least sensitive to the presence of sugar. It is suggested that hemagglutinins observed in plant pathogenic bacteria and those in plant host are similar and that both may, in some way, be involved in the plant-parasite relationship.

Hypersensitivity pneumonitis in grain farmers due to sensitization to Erwinia herbicola.

Two cases of hypersensitivity pneumonitis in grain workers are described. Both cases had evidence of sensitization to the gram-negative bacteria Erwinia herbicola as judged by skin tests, detection of serum precipitins, and inhalation challenge. E. herbicola is frequently found in the microflora of grains.